Candidate: Dr. Ghanem, M.D., is an Assistant Professor of Infectious Diseases and a Ph.D. candidate in the Graduate Training Program in Clinical Investigation at Johns Hopkins University School of Medicine. The goal of this K23 resubmission is to enhance his practical research skills and laboratory-based skills through focused didactics and the combined expertise of mentors and collaborators. These skills will complement the theoretical skills he has acquired thus far in his training, allowing him to become an independent and productive translational researcher. Background: Over 100 million women worldwide use hormonal contraceptives (HC). Their effects on mucosal and systemic immune responses are poorly understood. Observational studies have linked HC use to increased risk of sexually transmitted infection (STI) acquisition, including HIV. Preliminary Data: We have shown that cytokine production in a human-derived cervical and vaginal epithelial cell culture model was influenced by sex hormones in a compartment-specific manner. This finding was extended to a longitudinal study of 967 women where progesterone based HC were found to be independent predictors of lower genital tract inflammation. Specific Aims: 1) To compare cervicovaginal lavage fluid to endocervical, ectocervical, and vaginal wicks in evaluating lower genital tract mucosal immune responses; 2) To determine whether the use of HC affects cervicovaginal levels of soluble and cellular markers of inflammation in women infected with Neisseria gonorrhoeae, before and after therapy; and 3) To determine whether the use of HC affects systemic humoral and T-cell responses directed against N. gonorrhoeae outer membrane proteins. Significance: The findings from this proposal will help define optimal methods to measure genital immune responses; they will provide data that may guide the introduction of STI screening strategies specifically targeting HC users, and they will help define the potential role of sex hormones as adjuvants for mucosal vaccines in women.